PacBio RS II sequencing of SAMD00432597
For the vegetative cultures, mt+ (NIES-67) and mt- (NIES-68) cells were separately harvested at 6 (start time of light illumination is 0) and 18 h (IsoSeq 1). For the sexually-differentiated cells, vegetatively growing cells of the mt+ and mt- were separately collected, washed three times with MI medium, and incubated separately in MI medium (3.6E5 cells/ 72 mL in 300-mL Erlenmeyer flasks) under continuous light for 4 and 24 h (IsoSeq 2 and IsoSeq 3, respectively). For the mating-induced cells, cells of both mating types, which had been separately cultured in MI medium (at 3.6E5/ 72 mL in 300-mL Erlenmeyer flasks) for 24 h, were mixed and co-incubated (at 3.6E5 each/ 72 mL in 300-mL Erlenmeyer flasks) for 4 and 8 h (IsoSeq 4), 16 and 24 (IsoSeq 5), and 48 and 72 (IsoSeq 6). The harvested cells were frozen in liquid nitrogen and total RNA was isolated using TRIZOL Plus Kit. Six cDNA libraries (IsoSeq 1 to IsoSeq 6) were prepared using SMARTer PCR cDNA Synthesis kit (Clontech, www.takarabio.com), except that barcoded oligo-dT primers (dT-BC1 to dT-BC6, respectively) were used instead of 3' SMART CDS primer IIA included in the kit. These barcoded libraries were mixed and separated into 9 different insert sizes by SageELF (SAGE Science, www.sagescience.com). Eleven Libraries for Iso-Seq analysis were prepared using a SMRTbell Template Prep Kit v1.0 (Pacific Bioscience). Sequences of barcoded oligo-dT primers were AAGCAGTGGTATCAACGCAGAGTACtcagacgatgcgtcatTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTVN (dT_BC1),  AAGCAGTGGTATCAACGCAGAGTACctatacatgactctgcTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTVN  (dT_BC2), AAGCAGTGGTATCAACGCAGAGTACtactagagtagcactcTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTVN (dT_BC3), AAGCAGTGGTATCAACGCAGAGTACtgtgtatcagtacatgTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTVN (dT_BC4), AAGCAGTGGTATCAACGCAGAGTACgatctctactatatgcTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTVN (dT_BC5), and AAGCAGTGGTATCAACGCAGAGTACacagtctatactgctgTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTVN (dT_BC6).